Perfused rat hindlimb wound model: lambda-carrageenan induced

We established an isolated perfused rat hindlimb wound model using intramuscular injections of lambda-carrageenan. Wound and control hindlimbs of 24-h fasting anesthetized (phenobarbital sodium, 5 mg/100 g body wt) rats were perfused for 60 min (10 ml/min) with media containing 10 mM glucose and physiological levels of amino acids. Analysis of perfusate showed a significant increase in glucose clearance and lactate production from the wounded hindlimb but no significant difference in tissue adenine nucleotide levels. Alterations in net amino acid release were measured over a 60-min period; all amino acids were released in greater concentration from wounded hindlimbs. Comparison of amino acid release, normalized for phenylalanine concentration, suggested the following alterations in amino acid metabolism by the wounded tissue: depressed threonine, glycine serine and alanine release, and increased release of the branched chain amino acids (BCAA), leucine, isoleucine, and valine. The increased release and intracellular concentration of BCAA was supported by decreased BCAA transaminase activity in the wounded muscle tissue. This result may explain the decreased alanine release from the wounded hindlimbs.